PROJECT SUMMARY/ABSTRACT This grant proposal describes a five-year mentored training program for the career development of Dr. Jocelyn Kim, a physician-scientist, who will develop cutting-edge methods to quantify the HIV reservoir and investigate novel therapies to cure HIV infection. She has recently discovered that natural killer (NK) cells delays viral rebound in a sophisticated humanized mouse model of HIV latency. She has also found that anti-HIV chimeric antigen receptor (CAR)-on NK cells efficiently kill HIV-infected cells in vitro. In addition, a novel synthetic latency reactivating agent (LRA) activates latent HIV in the presence of ART in vivo. Importantly, she has innovated a genetically barcoded reporter HIV-1 and demonstrated that this diverse virus swarm is replication-competent and pathogenic in vivo and forms a latent reservoir, which rebounds after cessation of antiretroviral therapy (ART). She will use this novel barcoded-virus tool to accurately quantify the number of latently infected clones in vivo. These preliminary data are the basis of the current proposal, which involves the completion of the following aims. First, she will investigate whether NK cells from peripheral blood or hematopoietic stem cell progenitors (HSCPs) are more effective in delaying HIV rebound in vivo using new methods of stem cell differentiation and analysis. Second, she will test of efficacy of CAR-NK cells versus CAR-T cells on viral rebound in vivo. Lastly, she will use state-of-the-art approach in genetic barcoded virus to determine whether combinatorial therapies (LRA and NK cell-based therapies) affect the HIV reservoir in vivo. With K08 funding, she we will complete these studies with the intention of deepening our understanding of HIV pathogenesis and curative strategies. Dr. Kim is currently an assistant clinical professor in the UCLA Division of Infectious Diseases and recent graduate of the UCLA Specialty Training and Advanced Research (STAR) program, from which she received a PhD in Biology and Biological Engineering from California Institute of Technology under her PhD advisor Dr. David Baltimore. During the proposed award period, she will continue under the primary mentorship of Dr. Jerome Zack at UCLA with additional mentorship through an advisory committee comprised of Drs. Gay Crooks (UCLA), Alex Hoffman (UCLA) and Judith Currier (UCLA). With their guidance, she will attain the necessary research and career skills to apply successfully for independent funding and transition to independence. As such, through her collaborations as well as didactic training, she will expand her NK cell project and use complex computational skills for high-throughout barcode sequencing analysis to understand the HIV reservoir. She will also acquire new skills in stem cell differentiation and analysis, molecular cloning, and writing and grantsmanship. K08 funding will allow her to develop all of the skills and tools needed to become a successful independent investigator.